Inositol Pyrophosphate Profiling of Two HCT116 Cell Lines Uncovers Variation in InsP8 Levels
Fig 3
Comparisons of HCT116NIH and HCT116UCL cells: expression of IP6Ks and PPIP5Ks, capacity to dephosphorylate InsP8, cell growth, and phalloidin staining.
The following analyses of HCT116NIH and HCT116UCL cells were performed: Panel A, Western analyses of IP6Ks and PPIP5Ks. Complete gels, and procedures used to validate the antibodies, are described in S1 and S2 Figs. Panel B, quantitative RT-PCR analysis of expression of IP6K1, IP6K2 and IP6K3. Panel C, HPLC analysis of 1 μM [3H]InsP8 dephosphorylation by 70 μg cell lysates in 100 μl medium. Panel D, counting of cell growth for the indicated number of days. Panel E, labeling of the actin cytoskeleton with FITC-phalloidin. Hoechst was used as a nuclear stain.